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Updated: May 12, 2026
08:34
OaAEP1-Mediated Enzymatic Synthesis and Immobilization of Polymerized Protein for Single-Molecule Force Spectroscopy
Published on: February 5, 2020
Large scale modification of biomolecules using immobilized sortase A from Staphylococcus aureus
Max Steinhagen1, Katja Zunker, Karoline Nordsieck
1Institute of Biochemistry, Faculty of Biosciences, Pharmacy and Psychology, Universität Leipzig, Brüderstr. 34, 04103 Leipzig, Germany. msteinha@uni-leipzig.de
Bioorganic & Medicinal Chemistry
|April 20, 2013
Summary
Sortase A (SrtA) enzyme enables large-scale protein modification and biotinylation. Immobilizing SrtA on resin allows enzyme recycling and simplifies purification, accelerating bioprocesses.
Area of Science:
- Biochemistry
- Protein Engineering
- Biotechnology
Background:
- Sortase A (SrtA) from Staphylococcus aureus is recognized for its site-specific protein modification capabilities.
- SrtA has primarily been utilized for analytical-scale modifications to study biomolecules in cellular contexts.
Purpose of the Study:
- To demonstrate the applicability of SrtA-mediated ligation for large-scale protein modification.
- To optimize SrtA ligation for industrial applications and develop efficient purification strategies.
Main Methods:
- Optimization of SrtA-mediated ligation using peptides.
- Large-scale biotinylation of interleukin-8 under optimized conditions.
- C-terminal immobilization of SrtA onto a polyethylene glycol (PEG)-based resin.
Main Results:
- Successful optimization of SrtA ligation for large-scale applications.
- Demonstrated efficient, large-scale biotinylation of interleukin-8.
- Established stable, active, and recyclable immobilized SrtA.
- Facilitated purification processes by enabling centrifugation/filtration over chromatography.
Conclusions:
- SrtA-mediated ligation is applicable for large-scale site-specific protein modification.
- Immobilized SrtA offers significant advantages for bioprocessing, including enzyme recyclability and simplified purification.
- This approach accelerates protein modification workflows and enhances process efficiency.
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